鼻疽诺卡菌菌体蛋白的免疫蛋白质组学研究

摘要

Objective To identify the immunogenic proteins inNocardia farcinica cellular proteins and provide evidence for the screening of specific diagnostic antigens forN. farcinica. Methods The cellular proteins of N. farcinica IFM 10152 were extracted to perform two-dimensional polyacrylamide electrophoresis and western blot assay to screen immunogenic proteins which can react withN. farcinica infected rabbit sera. Results Through immunoblotting of the cellular proteins of N. farcinica, 9 proteins with antigenicity were found, in which 8 proteins had matching spots on 2-DE gel. After in-gel digestion, 8 immunogenic proteins were identified by MALDI - TOF MS, which were all in cytoplasm. Conclusion A immunoproteomics assay was established in this study by which novel immunogenic proteins were found inN. farcinica cellular proteins. This assay can be used in the screening of specific diagnostic antigens forN. farcinica in the research of diagnostic reagent of N. farcinica.%目的 通过鉴别鼻疽诺卡菌菌体蛋白中具有免疫原性的蛋白,为筛选鼻疽诺卡菌特异诊断抗原提供线索.方法提取鼻疽诺卡菌菌体蛋白进行双向电泳,结合免疫印迹法筛选与抗鼻疽诺卡菌兔血清发生免疫反应的蛋白点.结果 在鼻疽诺卡菌菌体蛋白的免疫杂交膜上,共发现9个具有抗原活性的蛋白,其中8个可在考马斯蓝染色胶上找到匹配点,通过介质辅助激光解吸电离飞行时间质谱成功鉴定8个蛋白,全部定位于胞内.结论 本研究成功建立鼻疽诺卡菌免疫蛋白质组学方法,在鼻疽诺卡菌菌体蛋白中发现新的具有免疫原性的蛋白,可作为鼻疽诺卡菌特异性诊断候选抗原,用于诊断试剂的研究.